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Crude protein extraction protocol for phage N15 protelomerase in vitro enzymatic assays
- Source :
- Analytical Biochemistry. 414:169-171
- Publication Year :
- 2011
- Publisher :
- Elsevier BV, 2011.
-
Abstract
- The phage N15 protelomerase enzyme (TelN) is essential for the replication of its genome by resolution of its telRL domain, located within a telomerase occupancy site (tos), into hairpin telomeres. Isolation of TelN for in vitro processing of tos, however, is a highly complex process, requiring multiple purification steps. In this study a simplified protocol for crude total protein extraction is described that retains the tos-cleaving activity of TelN for at least 4 weeks, greatly simplifying in vitro testing of its activity. This protocol may be extended for functional analysis of other phage and bacterial proteins, particularly DNA-processing enzymes.
Details
- ISSN :
- 00032697
- Volume :
- 414
- Database :
- OpenAIRE
- Journal :
- Analytical Biochemistry
- Accession number :
- edsair.doi.dedup.....da06bf439a7d0b02863a6fb2f2794b72